scholarly journals Effect of recombinant hematopoietic growth factors on proliferation of human marrow progenitor cells in serum-deprived liquid culture

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1387-1392 ◽  
Author(s):  
AR Migliaccio ◽  
G Migliaccio ◽  
JW Adamson
Keyword(s):  
Growth Factors ◽  
T Cell ◽  
Progenitor Cells ◽  
Liquid Culture ◽  
Adherent Cell ◽  
Hematopoietic Growth Factors ◽  
Gm Csf ◽  
Normal Human ◽  
Fetal Bovine ◽  
Marrow Cells

Abstract We investigated the effects of recombinant interleukin-3 (IL-3), granulocyte-macrophage and granulocyte colony-stimulating factors (GM- CSF and G-CSF), and erythropoietin (Ep) on the number of human hematopoietic progenitors after two to ten days of incubation in liquid cultures deprived of fetal bovine serum (FBS). The source of progenitor cells was normal human marrow depleted of T lymphocytes and/or adherent cells. When adherent cell-depleted marrow was cultured without growth factors, the number of progenitor cells was relatively constant for periods up to eight days. In contrast, a progressive decline in the number of progenitor cells was detected in cultures of nonadherent, T- cell-depleted marrow cells. In both cases, the addition of IL-3 increased by two- to fourfold over input the number of erythroid burst- forming cells (BFU-E) per culture. The number of BFU-E peaked either at day 4 or 8. G-CSF had no effect on the number of progenitor cells per culture. GM-CSF and Ep had no effect in cultures of nonadherent marrow cells but maintained the number of BFU-E in cultures of nonadherent, T- cell-depleted marrow cells. The addition of a neutralizing anti-GM-CSF monoclonal antibody, but not anti-IL-3 neutralizing antiserum, decreased the number of BFU-E in cultures of nonadherent marrow cells. None of the growth factors investigated enhanced the number of GM progenitors to the same degree as the number of BFU-E. However, in cultures of nonadherent, T-cell-depleted marrow cells, IL-3 and GM-CSF maintained the number of GM progenitors up to eight days. These results indicate that IL-3 alone is capable of increasing the number of BFU-E and of maintaining the number of GM progenitors in liquid culture, whereas GM-CSF and Ep are capable of maintaining, but not increasing, BFU-E in this system.

scholarly journals Effect of recombinant hematopoietic growth factors on proliferation of human marrow progenitor cells in serum-deprived liquid culture

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1387-1392
Author(s):  
AR Migliaccio ◽  
G Migliaccio ◽  
JW Adamson
Keyword(s):  
Growth Factors ◽  
T Cell ◽  
Progenitor Cells ◽  
Liquid Culture ◽  
Adherent Cell ◽  
Hematopoietic Growth Factors ◽  
Gm Csf ◽  
Normal Human ◽  
Fetal Bovine ◽  
Marrow Cells

We investigated the effects of recombinant interleukin-3 (IL-3), granulocyte-macrophage and granulocyte colony-stimulating factors (GM- CSF and G-CSF), and erythropoietin (Ep) on the number of human hematopoietic progenitors after two to ten days of incubation in liquid cultures deprived of fetal bovine serum (FBS). The source of progenitor cells was normal human marrow depleted of T lymphocytes and/or adherent cells. When adherent cell-depleted marrow was cultured without growth factors, the number of progenitor cells was relatively constant for periods up to eight days. In contrast, a progressive decline in the number of progenitor cells was detected in cultures of nonadherent, T- cell-depleted marrow cells. In both cases, the addition of IL-3 increased by two- to fourfold over input the number of erythroid burst- forming cells (BFU-E) per culture. The number of BFU-E peaked either at day 4 or 8. G-CSF had no effect on the number of progenitor cells per culture. GM-CSF and Ep had no effect in cultures of nonadherent marrow cells but maintained the number of BFU-E in cultures of nonadherent, T- cell-depleted marrow cells. The addition of a neutralizing anti-GM-CSF monoclonal antibody, but not anti-IL-3 neutralizing antiserum, decreased the number of BFU-E in cultures of nonadherent marrow cells. None of the growth factors investigated enhanced the number of GM progenitors to the same degree as the number of BFU-E. However, in cultures of nonadherent, T-cell-depleted marrow cells, IL-3 and GM-CSF maintained the number of GM progenitors up to eight days. These results indicate that IL-3 alone is capable of increasing the number of BFU-E and of maintaining the number of GM progenitors in liquid culture, whereas GM-CSF and Ep are capable of maintaining, but not increasing, BFU-E in this system.

scholarly journals Growth factor requirements of childhood acute T-lymphoblastic leukemia: correlation between presence of chromosomal abnormalities and ability to grow permanently in vitro

Blood ◽  
1991 ◽  
Vol 77 (7) ◽  
pp. 1534-1545 ◽  
Author(s):  
R O'Connor ◽  
A Cesano ◽  
B Lange ◽  
J Finan ◽  
PC Nowell ◽  
...  
Keyword(s):  
Growth Factors ◽  
T Cell ◽  
Growth Factor ◽  
Cell Lines ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Hematopoietic Growth Factors ◽  
Gm Csf

Cells from 10 cases of childhood acute T-lymphoblastic leukemia (T-ALL) were cultured in the presence of recombinant human interleukins (rhIL) or colony-stimulating factors (CSF) to analyze their growth factor requirements and differentiative potential. Although cells from most leukemic samples displayed a short-term proliferative response to several hematopoietic growth factors, only the ones featuring chromosomal translocations could be established as permanent cell lines. Two cell lines could be initiated only in the presence of IL-3 (TALL-103 and TALL-106), one in granulocyte-macrophage CSF (GM-CSF) (TALL-101), and one in IL-2 (TALL-104); only one cell line (TALL-105) was originated in the absence of growth factors. The TALL-101 and TALL- 103 cell lines, derived from very immature T-ALL cases, underwent growth factor-dependent phenotypic conversion (lymphoid to myeloid). However, the T-cell receptor rearrangement and karyotype of the original leukemic clones were retained. In contrast, the TALL-104, - 105, and -106 cell lines which originated from more mature T-ALL cases, maintained a T-lymphoblastic phenotype regardless of the growth factors in which they were expanded. These data demonstrate in vitro the aggressive nature of T-ALL cases bearing chromosomal abnormalities, and indicate that the lineage commitment of the malignant clone depends on its stage of maturation in T-cell ontogeny.

scholarly journals Growth factor requirements of childhood acute T-lymphoblastic leukemia: correlation between presence of chromosomal abnormalities and ability to grow permanently in vitro

Blood ◽  
1991 ◽  
Vol 77 (7) ◽  
pp. 1534-1545 ◽  
Author(s):  
R O'Connor ◽  
A Cesano ◽  
B Lange ◽  
J Finan ◽  
PC Nowell ◽  
...  
Keyword(s):  
Growth Factors ◽  
T Cell ◽  
Growth Factor ◽  
Cell Lines ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Hematopoietic Growth Factors ◽  
Gm Csf

Abstract Cells from 10 cases of childhood acute T-lymphoblastic leukemia (T-ALL) were cultured in the presence of recombinant human interleukins (rhIL) or colony-stimulating factors (CSF) to analyze their growth factor requirements and differentiative potential. Although cells from most leukemic samples displayed a short-term proliferative response to several hematopoietic growth factors, only the ones featuring chromosomal translocations could be established as permanent cell lines. Two cell lines could be initiated only in the presence of IL-3 (TALL-103 and TALL-106), one in granulocyte-macrophage CSF (GM-CSF) (TALL-101), and one in IL-2 (TALL-104); only one cell line (TALL-105) was originated in the absence of growth factors. The TALL-101 and TALL- 103 cell lines, derived from very immature T-ALL cases, underwent growth factor-dependent phenotypic conversion (lymphoid to myeloid). However, the T-cell receptor rearrangement and karyotype of the original leukemic clones were retained. In contrast, the TALL-104, - 105, and -106 cell lines which originated from more mature T-ALL cases, maintained a T-lymphoblastic phenotype regardless of the growth factors in which they were expanded. These data demonstrate in vitro the aggressive nature of T-ALL cases bearing chromosomal abnormalities, and indicate that the lineage commitment of the malignant clone depends on its stage of maturation in T-cell ontogeny.

scholarly journals Interleukin 6 is a permissive factor for monocytic colony formation by human hematopoietic progenitor cells.

1992 ◽  
Vol 175 (4) ◽  
pp. 1151-1154 ◽  
Author(s):  
J H Jansen ◽  
J C Kluin-Nelemans ◽  
J Van Damme ◽  
G J Wientjens ◽  
R Willemze ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Progenitor Cells ◽  
Interleukin 6 ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Gm Csf ◽  
Marrow Cells

Since monocytes and macrophages that arise during the culture of bone marrow progenitor cells are potential sources of interleukin 6 (IL-6), we investigated whether auto- or paracrine production of this factor is involved in colony formation by normal hematopoietic progenitor cells. We added a polyclonal anti-IL-6 antiserum and a monoclonal anti-IL-6 antibody to cultures of monocyte- and T cell-depleted bone marrow cells. Colony formation was stimulated with granulocyte/monocyte-colony-stimulating factor (GM-CSF), monocyte-CSF, or IL-3. Addition of anti-IL-6 antibody resulted in decreased numbers of monocytic colonies to 40-50% of control values, whereas the numbers of granulocytic colonies were not altered. The inhibitory effect was preserved in cultures of CD34(+)-enriched bone marrow cells. As a second approach, we added a monoclonal antibody directed against the IL-6 receptor to cultures of monocyte- and T cell-depleted bone marrow cells. This antibody almost completely inhibited the growth of monocytic colonies, again without decreasing the number of granulocytic colonies. Finally, the importance of IL-6 in monocytopoiesis was demonstrated in serum-deprived bone marrow cultures: addition of exogenous IL-6 to cultures stimulated with GM-CSF resulted in increased numbers of monocytic colonies. Our results indicate that the permissive presence of IL-6 is required for optimal monocytic colony formation by bone marrow progenitor cells.

scholarly journals Activation of adenosine A3 receptors potentiates stimulatory effects of IL-3, SCF, and GM-CSF on mouse granulocyte-macrophage hematopoietic progenitor cells

2009 ◽  
pp. 247-252
Author(s):  
M Hofer ◽  
A Vacek ◽  
M Pospíšil ◽  
J Holá ◽  
D Štreitová ◽  
...  
Keyword(s):  
Growth Factors ◽  
Progenitor Cells ◽  
Bone Marrow Cells ◽  
Hematopoietic Progenitor Cells ◽  
Mouse Bone Marrow ◽  
Colony Stimulating Factor ◽  
Hematopoietic Growth Factors ◽  
Hematopoietic Progenitor ◽  
Gm Csf ◽  
Stimulating Factor

Adenosine A3 receptor agonist N6-(3-iodobenzyl)adenosine-5’-Nmethyluronamide (IB-MECA) has been tested from the point of view of potentiating the effects of hematopoietic growth factors interleukin-3 (IL-3), stem cell factor (SCF), granulocytemacrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) on the growth of hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in suspension of normal mouse bone marrow cells in vitro. IB-MECA alone induced no GM-CFC growth. Significant elevation of numbers of GM-CFC evoked by the combinations of IB-MECA with IL-3, SCF, or GM-CSF as compared with these growth factors alone has been noted. Combination of IB-MECA with G-CSF did not induce significantly higher numbers of GM-CFC in comparison with G-CSF alone. Joint action of three drugs, namely of IB-MECA + IL-3 + GM-CSF, produced significantly higher numbers of GM-CFC in comparison with the combinations of IB-MECA + IL-3, IB-MECA + GM-CSF, or IL-3 + GM-CSF. These results give evidence of a significant role of selective activation of adenosine A3 receptors in stimulation of the growth of granulocyte/ macrophage hematopoietic progenitor cells.

Expansion of Bone Marrow (BM) Progenitors and Mobilization of Multilineage Progenitor Cells Associated with Once Per Cycle Administration of Long-Acting Growth Factors Neulasta and Aranesp with Chemotherapy (CT): Biology and Nature of Response.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2905-2905
Author(s):  
Saroj Vadhan-Raj ◽  
Carlos E. Bueso-Ramos ◽  
G. Hangoc ◽  
K. Christopherson ◽  
M. Collard ◽  
...  
Keyword(s):  
Growth Factors ◽  
Progenitor Cells ◽  
Receptor Activation ◽  
High Dose ◽  
Hematopoietic Growth Factors ◽  
Gm Csf ◽  
Kit Ligand ◽  
Cd 34 ◽  
Biologic Effects ◽  
Long Acting

Abstract Hematopoietic growth factors (HGF) G-CSF and GM-CSF have been utilized widely to facilitate mobilization of progenitor cells that can be used to support high dose CT. Recently long-acting HGF neulasta (pegfilgrastim) and aranesp (darbepoetin alfa) were developed to reduce the frequency of administration. While these agents are effective in reducing CT-induced neutropenia and anemia similar to their parent compounds, very little is known about their efficiency in mobilizing progenitor cells. The purpose of this study was to evaluate biologic effects of these agents, used in combination with CT, on progenitor cells. Chemo-naïve sarcoma patients receiving adriamycin and ifosfamide (AI) were treated once per cycle with aranesp (500 mcg) SC prior to initiating CT (day 0) and neulasta (6 mg) SC after completion of CT (day 4). BM and peripheral blood (PB) samples were studied at the baseline and around day 14 (at the time of WBC recovery) for progenitors and mediators of response. The treatment was associated with a significant increase in PB CD 34 + cells (median increase 36-fold, p=0.005) and marked mobilization (p=0.003) of CFU-GM (24-fold), BFU-E (22-fold), and CFU-GEMM (62-fold) (n=14). To better understand the biology and nature of response, we examined BM before and at the time of mobilization (n=12). There was an expansion of multi-lineage BM progenitors (p=0.06) and CD 34+ cells (P=0.001). BM exam at the time of recovery showed increased cellularity (2-fold) with increased granulopoietic elements. An increased expression of MMP-9 but unchanged expression of TIMP-1 was observed by Immunohistochemistry (IHC). c-kit ligand level by Elisa was decreased in the BM supernatant. In addition, phospho c-kit (phospho site 568) was increased in the myeloid and erythroid precursors by IHC but phospho site 823 was unaltered, indicating the specificity of activation of the downstream pathway in response to c-kit ligand-receptor activation, possibly leading to expansion of progenitor cells. Interestingly, expression of CXCR4 at the protein level (flow cytometry and western blot) and RNA level (RT-PCR) was decreased (p<0.05) in the BM at the time of mobilization (day 14) as compared to the baseline. These findings indicate that long-acting HGFs, administered once per cycle of CT, are a potent stimulus for mobilization of myeloid, erythroid, and multipotential progenitors. Our findings also suggest that increased myelopiesis in the regenerating BM may be associated with increased protease activity such as MMP-9 that may result in the cleavage of c-kit leading to activation of c-kit receptors and expansion of progenitors, and the cleavage of CXCR-4 leading to release of progenitors from the BM anchorage and mobilization into PB.

scholarly journals Developmental regulation of erythropoiesis by hematopoietic growth factors: analysis on populations of BFU-E from bone marrow, peripheral blood, and fetal liver

Blood ◽  
1989 ◽  
Vol 74 (1) ◽  
pp. 49-55 ◽  
Author(s):  
SG Emerson ◽  
S Thomas ◽  
JL Ferrara ◽  
JL Greenstein
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Fetal Liver ◽  
Developmental Regulation ◽  
Hematopoietic Growth Factors ◽  
Erythroid Progenitor ◽  
Gm Csf ◽  
Proliferative Effect

Abstract Fetal hematopoiesis is characterized by expanding erythropoiesis to support a continuously increasing RBC mass. To explore the basis for this anabolic, nonhomeostatic erythropoiesis, the proliferative effect of recombinant hematopoietic growth factors on highly enriched hematopoietic progenitor cells from fetal and adult tissues were compared. Fetal hepatic BFU-E, unlike adult bone marrow (BM) or peripheral blood (PB) BFU-E, were capable of proliferating in response to erythropoietin in the absence of added GM colony-stimulating factor (GM-CSF) or interleukin-3 (IL-3), and erythropoietin (Epo) directly stimulated the expansion of the fetal BFU-E pool in suspension culture. A murine monoclonal antibody (MoAb), Ep 3, was raised against enriched fetal liver progenitor cells, which detected all fetal BFU-E and which reacted with the erythropoietin-responsive, GM-CSF/IL-3-independent fraction of adult BM BFU-E and CFU-E. All adult PB BFU-E were Ep 3- but became Ep 3+ after stimulation with GM-CSF or IL-3. These data indicate that Epo plays a unique role in fetal hepatic erythropoiesis, stimulating proliferation of immature BFU-E in addition to promoting terminal differentiation of later erythroid progenitor cells. In addition, these results demonstrate a MoAb which detects all erythropoietin-responsive progenitor cells and distinguishes the BFU-E compartments in adult BM and PB.

scholarly journals Developmental regulation of erythropoiesis by hematopoietic growth factors: analysis on populations of BFU-E from bone marrow, peripheral blood, and fetal liver

Blood ◽  
1989 ◽  
Vol 74 (1) ◽  
pp. 49-55
Author(s):  
SG Emerson ◽  
S Thomas ◽  
JL Ferrara ◽  
JL Greenstein
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Fetal Liver ◽  
Developmental Regulation ◽  
Hematopoietic Growth Factors ◽  
Erythroid Progenitor ◽  
Gm Csf ◽  
Proliferative Effect

Fetal hematopoiesis is characterized by expanding erythropoiesis to support a continuously increasing RBC mass. To explore the basis for this anabolic, nonhomeostatic erythropoiesis, the proliferative effect of recombinant hematopoietic growth factors on highly enriched hematopoietic progenitor cells from fetal and adult tissues were compared. Fetal hepatic BFU-E, unlike adult bone marrow (BM) or peripheral blood (PB) BFU-E, were capable of proliferating in response to erythropoietin in the absence of added GM colony-stimulating factor (GM-CSF) or interleukin-3 (IL-3), and erythropoietin (Epo) directly stimulated the expansion of the fetal BFU-E pool in suspension culture. A murine monoclonal antibody (MoAb), Ep 3, was raised against enriched fetal liver progenitor cells, which detected all fetal BFU-E and which reacted with the erythropoietin-responsive, GM-CSF/IL-3-independent fraction of adult BM BFU-E and CFU-E. All adult PB BFU-E were Ep 3- but became Ep 3+ after stimulation with GM-CSF or IL-3. These data indicate that Epo plays a unique role in fetal hepatic erythropoiesis, stimulating proliferation of immature BFU-E in addition to promoting terminal differentiation of later erythroid progenitor cells. In addition, these results demonstrate a MoAb which detects all erythropoietin-responsive progenitor cells and distinguishes the BFU-E compartments in adult BM and PB.

scholarly journals Ex vivo expansion of human hematopoietic stem and progenitor cells

Blood ◽  
2011 ◽  
Vol 117 (23) ◽  
pp. 6083-6090 ◽  
Author(s):  
Ann Dahlberg ◽  
Colleen Delaney ◽  
Irwin D. Bernstein
Keyword(s):  
Stem Cell ◽  
Growth Factors ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Hematopoietic Growth Factors ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Stem And Progenitor Cells

AbstractDespite progress in our understanding of the growth factors that support the progressive maturation of the various cell lineages of the hematopoietic system, less is known about factors that govern the self-renewal of hematopoietic stem and progenitor cells (HSPCs), and our ability to expand human HSPC numbers ex vivo remains limited. Interest in stem cell expansion has been heightened by the increasing importance of HSCs in the treatment of both malignant and nonmalignant diseases, as well as their use in gene therapy. To date, most attempts to ex vivo expand HSPCs have used hematopoietic growth factors but have not achieved clinically relevant effects. More recent approaches, including our studies in which activation of the Notch signaling pathway has enabled a clinically relevant ex vivo expansion of HSPCs, have led to renewed interest in this arena. Here we briefly review early attempts at ex vivo expansion by cytokine stimulation followed by an examination of our studies investigating the role of Notch signaling in HSPC self-renewal. We will also review other recently developed approaches for ex vivo expansion, primarily focused on the more extensively studied cord blood–derived stem cell. Finally, we discuss some of the challenges still facing this field.

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